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At least 991 records · Page 55Linked to original sources
PubMed2006

Application of two-dimensional gel electrophoresis to interrogate alterations in the proteome of genetically modified crops. 1. Assessing analytical validation.

Current tools used to assess the safety of food and feed derived from modern biotechnology emphasize the investigation of possible unintended effects caused directly by the expression of transgenes or indirectly by pleiotropy. These tools include extensive multisite and multiyear agronomic evaluations, compositional analyses, animal nutrition, and classical toxicology evaluations. Because analytical technologies are rapidly developing, proteome analysis based on two-dimensional gel electrophoresis (2DE) was investigated as a complementary tool to the existing technologies. A 2DE method was established for the qualitative and quantitative analysis of the seed proteome of Arabidopsis thaliana with the following validation parameters examined: (1) source and scope of variation; (2) repeatability; (3) sensitivity; and (4) linearity of the method. The 2DE method resolves proteins with isoelectric points between 4 and 9 and molecular masses (MM) of 6-120 kDa and is sensitive enough to detect protein levels in the low nanogram range. The separation of the proteins was demonstrated to be very reliable with relative position variations of 1.7 and 1.1% for the pI and MM directions, respectively. The mean coefficient of variation of 254 matched spot qualities was found to be 24.8% for the gel-to-gel and 26% for the overall variability. A linear relationship (R2 > 0.9) between protein amount and spot volume was demonstrated over a 100-fold range for the majority of selected proteins. Therefore, this method could be used to interrogate proteome alterations such as a novel protein, fusion protein, or any other change that affects molecular mass, isoelectric point, and/or quantity of a protein.

Arabidopsis↗
PubMed2006

Evaluation of a non-targeted "omic" approach in the safety assessment of genetically modified plants.

Genetically modified plants must be approved before release in the European Union, and the approval is generally based upon a comparison of various characteristics between the transgenic plant and a conventional counterpart. As a case study, focusing on safety assessment of genetically modified plants, we here report the development and characterisation of six independently transformed ARABIDOPSIS THALIANA lines modified in the flavonoid biosynthesis. Analyses of integration events and comparative analysis for characterisation of the intended effects were performed by PCR, quantitative Real-time PCR, and High Performance Liquid Chromatography. Analysis by cDNA microarray was used as a non-targeted approach for the identification of potential unintended effects caused by the transformation. The results revealed that, although the transgenic lines possessed different types of integration events, no unintended effects were identified. However, we found that the majority of genes showing differential expression were identified as stress-related genes and that environmental conditions had a large impact on the expression of several genes, proteins, and metabolites. We suggest that the microarray approach has the potential to become a useful tool for screening of unintended effects, but state that it is crucial to have substantial information on the natural variation in traditional crops in order to be able to interpret "omics" data correctly within the framework of food safety assessment strategies of novel plant varieties, including genetically modified plant varieties.

Arabidopsis↗
PubMed2001

Quality of Life Programme--food, nutrition, and health--projects promotion.

The EC Quality of Life Programme (QoL), Key Action 1--Food, Nutrition & Health aims at providing a healthy, safe, and high-quality food supply leading to reinforced consumer's confidence in the safety of the European food. Key Action 1 is currently supporting several European projects investigating analytical methods for food control including sensors, risk analysis, and food safety standardisation. Their objectives range from the development and validation of prevention strategies for mycotoxin formation via the development of a communication platform for Genetically Modified Organisms (GMO), validation and standardisation of diagnostic Polymerase Chain Reaction (PCR) for food-borne pathogens, up to the evaluation of the potential cancer-preventing activity of pro- and pre-biotic ("SYNBIOTIC") combinations in human volunteers. This paper also informs on future research needs in food safety.

Consumer Product Safety↗
PubMed2004

Crop protection--what will shape the future picture?

The drivers which influence the types of crop protection most needed by agriculture are changing. A polarisation of approaches has resulted in the needs of organic agriculture and biotechnological agriculture, which can be identified as the current extremes of the spectrum, becoming very different. The main requirements of these two sectors are identified here and used as the basis for questioning future EU research requirements in crop protection. Factors affecting/influencing organic farming and the use of genetically modified crops are discussed in some detail.

Crops, Agricultural↗
PubMed2001

Food allergy: what are the issues?

With a growing interest in the development of genetically modified crop plants there is a need for appropriate approaches to safety assessment. Among the issues that have to be addressed is consideration of whether the products of novel genes have the potential to cause allergic sensitization. Resulting from a collaboration between the International Food Biotechnology Council and the International Life Sciences Institute recommendations have been made for a step-wise approach to the assessment of allergenic potential based upon considerations of serological identity, and sequence or structural homology, with known allergens and examination of the stability of the test protein in a simulated gastric fluid. In parallel there has been interest in the development of animal models, which would permit a more direct evaluation of potential allergenic activity. Progress in these areas is reviewed briefly in the context of what is known of food allergy and some of the important issues, which must be addressed in designing safety assessment strategies identified.

Allergens↗
PubMed2001

A multiplex PCR method of detecting recombinant DNAs from five lines of genetically modified maize.

Seven lines of genetically modified (GM) maize have been authorized in Japan as foods and feeds imported from the USA. We improved a multiplex PCR method described in the previous report in order to distinguish the five lines of GM maize. Genomic DNA was extracted from GM maize with a silica spin column kit, which could reduce experimental time and improve safety in the laboratory and potentially in the environment. We sequenced recombinant DNA (r-DNA) introduced into GM maize, and re-designed new primer pairs to increase the specificity of PCR to distinguish five lines of GM maize by multiplex PCR. A primer pair for the maize intrinsic zein gene (Ze1) was also designed to confirm the presence of amplifiable maize DNA. The lengths of PCR products using these six primer pairs were different. The Ze1 and the r-DNAs from the five lines of GM maize were qualitatively detected in one tube. The specific PCR bands were distinguishable from each other on the basis of the expected length. The r-DNA could be detected from maize samples containing 0.5% of each of the five lines of GM maize. The sensitivity would be acceptable to secure the verification of non-GMO materials and to monitor the reliability of the labeling system.

DNA Primers↗
PubMedDate not supplied

Determination of Cry9C protein in processed foods made with StarLink corn.

StarLink (Aventis CropScience US) hybrid corn has been genetically modified to contain a pesticidal protein, Cry9C, which makes it more resistant than traditional varieties to certain types of corn insect pests. Unlike other varieties of genetically engineered corn, the U.S. Environmental Protection Agency authorized the use of StarLink corn for animal feed and industrial use only, not for human consumption. However, some Cry9C-containing corn was mistakenly or inadvertently comingled with yellow corn intended for human food use. Because corn containing the Cry9C construct was not approved for human use, the U.S. Food and Drug Administration considers food containing it to be adulterated. Consequently, this regulatory violation resulted in hundreds of recalls of corn-based products, such as taco shells, containing cry9C DNA. Detecting the novel protein in StarLink corn is an emerging issue; therefore, there is no standardized or established analytical method for detecting Cry9C protein in processed foods. We developed a procedure for quantitation of Cry9C protein, with validation data, in processed food matrixes with a limit of quantitation at 1.7 ng/g (ppb), using a commercial polyclonal antibody-based Cry9C kit that was intended for corn grain samples. Intra- and interassay coefficients of variation were 2.8 and 11.8%, respectively. Mean recoveries were 73 and 85% at 2 and 5 ng/g Cry9C fortifications, respectively, for 19 control non-StarLink corn-based matrixes. Our data demonstrate only 0-0.5% of Cry9C protein survived the processing of tortilla chips and soft tortillas made from 100% StarLink corn, resulting in levels from below the detection limit to 45 ppb.

Bacillus thuringiensis Toxins↗
PubMed1998

Development and application of a selective detection method for genetically modified soy and soy-derived products.

A method has been developed to distinguish between traditional soy beans and transgenic Roundup Ready soy beans, i.e. the glyphosate ('Roundup') resistant soy bean variety developed by Monsanto Company. Glyphosate resistance results from the incorporation of an Agrobacterium-derived 5-enol-pyruvyl-shikimate-3-phosphatesynthase (EPSPS) gene. The detection method developed is based on a nested Polymerase Chain Reaction (PCR) procedure. Ten femtograms of soy bean DNA can be detected, while, starting from whole soy beans, Roundup Ready DNA can be detected at a level of 1 Roundup Ready soy bean in 5000 non-GM soy beans (0.02% Roundup Ready soy bean). The method has been applied to samples of soy bean, soy-meal pellets and soy bean flour, as well as a number of processed complex products such as infant formula based on soy, tofu, tempeh, soy-based desserts, bakery products and complex meat and meat-replacing products. The results obtained are discussed with respect to practical application of the detection method developed.

DNA, Plant↗
PubMedDate not supplied

A decade of European field trials with genetically modified plants.

This article analyzes the development of notifications of genetically modified plants field trials in the European Union from 1991 to 2001, based on the data collected at the European level in the Summary Notification Information Format database. During this time period, a total of 1687 field trial notifications were received. The number of field trial notifications dropped by 76% between 1998 and 2001, mainly due to the de facto moratorium in place since 1999. Input traits (77%) dominated the field trial notifications during the last decade, while output traits were relevant in only 18% of all notifications, with a decreasing relevance during the last six years. In particular, field trial notifications on molecular farming were almost absent in the EU. Large companies focused their field trials on crops with a high grown area in the European Union and resistance traits, while public institutions showed interest in a large diversity of plants and traits. Finally, some conclusions on future impacts of the results of the study are drawn in this article.

Agriculture↗
PubMed2003

Responses of plants and invertebrate trophic groups to contrasting herbicide regimes in the Farm Scale Evaluations of genetically modified herbicide-tolerant crops.

Effects of genetically modified herbicide-tolerant (GMHT) and conventional crop management on invertebrate trophic groups (herbivores, detritivores, pollinators, predators and parasitoids) were compared in beet, maize and spring oilseed rape sites throughout the UK. These trophic groups were influenced by season, crop species and GMHT management. Many groups increased twofold to fivefold in abundance between early and late summer, and differed up to 10-fold between crop species. GMHT management superimposed relatively small (less than twofold), but consistent, shifts in plant and insect abundance, the extent and direction of these effects being dependent on the relative efficacies of comparable conventional herbicide regimes. In general, the biomass of weeds was reduced under GMHT management in beet and spring oilseed rape and increased in maize compared with conventional treatments. This change in resource availability had knock-on effects on higher trophic levels except in spring oilseed rape where herbivore resource was greatest. Herbivores, pollinators and natural enemies changed in abundance in the same directions as their resources, and detritivores increased in abundance under GMHT management across all crops. The result of the later herbicide application in GMHT treatments was a shift in resource from the herbivore food web to the detritivore food web. The Farm Scale Evaluations have demonstrated over 3 years and throughout the UK that herbivores, detritivores and many of their predators and parasitoids in arable systems are sensitive to the changes in weed communities that result from the introduction of new herbicide regimes.

Agriculture↗
PubMed2006

Human obesity: its hormonal basis and the role of gastric inhibitory polypeptide.

Obesity is an abnormal expansion of the adipose organ and is a pathophysiological response to an imbalance between energy intake and energy expenditure. It is the result of a large number of diverse factors involving heritable and environmental characteristics. A simple definition of obesity is difficult and unsatisfactory and its age dependency has largely been ignored. Differentiation between healthy, age-related plumpness and obesity is often blurred and responsible for overdiagnosis of obesity in the developed world. In the past, epidemiological studies have often ignored the different prognostic significance of the two major phenotypes of human obesity making their conclusions of limited value. The role of heritable factors in determining both the propensity to develop obesity under favourable environmental conditions, including inactivity and unlimited access to fat-rich foods, and the phenotype it assumes received an enormous fillip from experiments involving genetically modified animals. The most important of these have demonstrated the key role played by a number of newly discovered or recently resurrected polypeptide hormones that are released from the intestine in response to food. Molecular manipulation of these hormones, especially of glucose-dependent insulin-stimulatory polypeptide offers a new therapeutic approach.

Adipose Tissue↗